Abstract
ADP-ribosylation factor 6 small GTPase plays an important role in cell migration, invasion and angiogenesis, which are the hallmarks of cancer. Although alterations in ARF6 expression and activity have been linked to metastatic cancer in one or two tissues, the expression of ARF6 in cancers over a wide range of tissues has not been studied so far. In this report, we analysed the expression of ARF6 mRNA in cancers and corresponding healthy controls from 17 different tissues by real-time qualitative polymerase chain reaction (RT-qPCR). We further evaluated ARF6 protein expression in oesophageal adenocarcinoma (EAC) tissue microarray cores by immunohistochemistry. The ARF6 gene expression levels are highly variable between healthy and cancer tissues. Our findings suggest that the ARF6 gene expression is up-regulated highest in oesophageal cancer. In EAC TMAs, ARF6 protein expression increase correlated with EAC progression. This is the first study to investigate ARF6 gene expression in a wide array of cancer tissues and demonstrate that ARF6 expression, at both mRNA and protein levels, is significantly upregulated in higher grades of EAC, which may be useful in targeting ARF6 for cancer diagnostic and therapeutic purposes.
Highlights
Cancer is one of the leading causes of death in the world
The Polymerase chain reaction (PCR) products were run on a 1.5% agarose gel, which was prepared in Tris Acetate EDTA (TAE) containing 1μg/ml ethidium bromide, at 100 volts in TAE buffer for 30min, and the gel was visualised using a GelDoc (BioRad)
A standard curve plotted from the results showed that assay runs linearly across the whole dilution series and as little as 7.5fg (10 copies) of plasmid DNA per reaction could still be detected by the real-time qualitative polymerase chain reaction (RT-qPCR)
Summary
Cancer is one of the leading causes of death in the world. In 2018, there were an estimated 18.1 million new cases and 9.6 million cancer-related deaths worldwide [1]. Despite advances in early detection and effective treatments, globally cancer incidents are predicted to increase to >23 million annually by 2030 and its mortality rate has risen by 25% since the 1990s. The increase in cancer mortality rate and incidents is due to a possible combination of various factors such as exposure to carcinogens, unhealthy lifestyle choices, age, inflammation and a genetic predisposition from acquired or inherited polymorphisms [2,3,4]. Cancer is defined as an uncontrolled growth that is resistant to antigrowth signals, evades apoptosis, has unlimited replicative potential, can stimulate angiogenesis, and has invasion/
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